Wilkinson M J, Cohen R L, Shuman M A
Cancer Research Institute, University of California, San Francisco 94143.
Anal Biochem. 1990 Mar;185(2):294-6. doi: 10.1016/0003-2697(90)90295-k.
A sensitive assay for type IV collagen degradation using an avidin-biotin sandwich technique is described. Biotinylated type IV collagen is allowed to bind to an avidin-coated microtiter plate. The solution to be assayed is incubated with the biotinylated collagen bound to the avidin plate. Collagen degraded by the solution is released into the supernatant and transferred to a second plate coated with avidin. By addition of biotinylated horseradish peroxidase to this second plate, the amount of collagen degraded is determined. Our assay requires only 0.5 microgram of type IV collagen per microtiter plate and detects nanogram quantities of bacterial collagenase activity.
本文描述了一种使用抗生物素蛋白-生物素夹心技术检测IV型胶原降解的灵敏检测方法。将生物素化的IV型胶原与包被有抗生物素蛋白的微量滴定板结合。将待测溶液与结合在抗生物素蛋白板上的生物素化胶原一起孵育。被溶液降解的胶原释放到上清液中，并转移到另一块包被有抗生物素蛋白的板上。通过向第二块板中加入生物素化的辣根过氧化物酶，可测定降解的胶原量。我们的检测方法每块微量滴定板仅需0.5微克IV型胶原，可检测纳克量的细菌胶原酶活性。
